← Product Code [DJG](/submissions/TX/subpart-d%E2%80%94clinical-toxicology-test-systems/DJG) · K231752

# ARK Hydrocodone Assay (K231752)

_Ark Diagnostics, Inc. · DJG · Nov 9, 2023 · Clinical Toxicology · SESE_

**Canonical URL:** https://fda.innolitics.com/submissions/CH/subpart-d%E2%80%94clinical-toxicology-test-systems/DJG/K231752

## Device Facts

- **Applicant:** Ark Diagnostics, Inc.
- **Product Code:** [DJG](/submissions/TX/subpart-d%E2%80%94clinical-toxicology-test-systems/DJG.md)
- **Decision Date:** Nov 9, 2023
- **Decision:** SESE
- **Submission Type:** Traditional
- **Regulation:** 21 CFR 862.3650
- **Device Class:** Class 2
- **Review Panel:** Clinical Toxicology

## Indications for Use

The ARK Hydrocodone Assay is an immunoassay intended for the qualitative detection and/or semi-quantitative estimation of hydrocodone and its metabolites in human urine at a cutoff of 300 ng/mL. The semi-quantitative mode is for the purpose of (1) enabling laboratories to determine an appropriate dilution of the specimen for confirmation by a confirmatory method, such as Gas Chromatography/Mass Spectrometry (GC/MS) or Liquid Chromatography/tandem Mass Spectrometry (LC-MS/MS), or (2) permitting laboratories to establish quality control procedures. The ARK Hydrocodone Assay provides only a preliminary analytical test result. A more specific alternative chemical method must be used in order to obtain a confirmed positive analytical result. Gas Chromatography/Mass Spectrometry (GC/MS) or Liquid Chromatography/tandem Mass Spectrometry (LC-MS/MS) is the preferred confirmatory method. Clinical consideration and professional judgment should be exercised with any drug test result, particularly when the preliminary test result is positive.

## Device Story

ARK Hydrocodone Assay is a liquid, ready-to-use homogeneous enzyme immunoassay (HEIA) for human urine. It utilizes a competitive binding mechanism between hydrocodone in the specimen and hydrocodone labeled with recombinant glucose-6-phosphate dehydrogenase (rG6PDH) for antibody binding sites. Increased hydrocodone concentration in the sample leads to increased enzyme activity, which converts NAD to NADH; this change is measured spectrophotometrically at 340 nm. The device is used in clinical laboratories on automated chemistry analyzers by trained personnel. It provides preliminary analytical results requiring confirmation by GC/MS or LC-MS/MS. The assay aids clinicians in identifying hydrocodone presence, supporting clinical decision-making regarding patient drug use.

## Clinical Evidence

Bench testing only. Precision study (N=160/level) evaluated qualitative and semi-quantitative performance across 0-200% of cutoff. Linearity demonstrated from 0-800 ng/mL. Analytical specificity tested against structurally related opiates and non-related compounds; no significant interference observed. Method comparison study (N=226) against LC-MS/MS reference method showed acceptable agreement. No clinical studies performed.

## Technological Characteristics

Homogeneous enzyme immunoassay (HEIA). Two-reagent system: R1 (rabbit monoclonal anti-hydrocodone antibodies + substrate) and R2 (hydrocodone derivative labeled with bacterial recombinant G6PDH). Liquid, ready-to-use. Spectrophotometric detection at 340 nm. Automated clinical chemistry analyzer platform. Storage at 2-8°C. Sodium azide preservative.

## Regulatory Identification

An opiate test system is a device intended to measure any of the addictive narcotic pain-relieving opiate drugs in blood, serum, urine, gastric contents, and saliva. An opiate is any natural or synthetic drug that has morphine-like pharmocological actions. The opiates include drugs such as morphine, morphine glucoronide, heroin, codeine, nalorphine, and meperedine. Measurements obtained by this device are used in the diagnosis and treatment of opiate use or overdose and in monitoring the levels of opiate administration to ensure appropriate therapy.

## Special Controls

*Classification.* Class II (special controls). An opiate test system is not exempt if it is intended for any use other than employment or insurance testing or is intended for Federal drug testing programs. The device is exempt from the premarket notification procedures in subpart E of part 807 of this chapter subject to the limitations in § 862.9, provided the test system is intended for employment and insurance testing and includes a statement in the labeling that the device is intended solely for use in employment and insurance testing, and does not include devices intended for Federal drug testing programs (*e.g.,* programs run by the Substance Abuse and Mental Health Services Administration (SAMHSA), the Department of Transportation (DOT), and the U.S. military).

## Predicate Devices

- DRI Hydrocodone Assay ([K150502](/device/K150502.md))

## Submission Summary (Full Text)

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>
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FDA

U.S. FOOD & DRUG

ADMINISTRATION

# 510(k) SUBSTANTIAL EQUIVALENCE DETERMINATION DECISION SUMMARY

ASSAY ONLY

## I Background Information:

A 510(k) Number

K231752

B Applicant

ARK Diagnostics, Inc.

C Proprietary and Established Names

ARK Hydrocodone Assay

D Regulatory Information

|  Product Code(s) | Classification | Regulation Section | Panel  |
| --- | --- | --- | --- |
|  DJG | Class II | 21 CFR 862.3650 - Opiate Test System | TX - Clinical Toxicology  |

## II Submission/Device Overview:

A Purpose for Submission:

New Device

B Measurand:

Hydrocodone

C Type of Test:

Qualitative and semi-quantitative homogeneous immunoassay

## III Intended Use/Indications for Use:

A Intended Use(s):

See Indications for Use below.

Food and Drug Administration

10903 New Hampshire Avenue

Silver Spring, MD 20993-0002

www.fda.gov

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K231752 - Page 2 of 15

## B Indication(s) for Use:

The ARK Hydrocodone Assay is an immunoassay intended for the qualitative detection and/or semi-quantitative estimation of hydrocodone and its metabolites in human urine at a cutoff of 300 ng/mL. The semi-quantitative mode is for the purpose of (1) enabling laboratories to determine an appropriate dilution of the specimen for confirmation by a confirmatory method, such as Gas Chromatography/Mass Spectrometry (GC/MS) or Liquid Chromatography/tandem Mass Spectrometry (LC-MS/MS), or (2) permitting laboratories to establish quality control procedures.

The ARK Hydrocodone Assay provides only a preliminary analytical test result. A more specific alternative chemical method must be used in order to obtain a confirmed positive analytical result. Gas Chromatography/Mass Spectrometry (GC/MS) or Liquid Chromatography/tandem Mass Spectrometry (LC-MS/MS) is the preferred confirmatory method. Clinical consideration and professional judgment should be exercised with any drug test result, particularly when the preliminary test result is positive.

## C Special Conditions for Use Statement(s):

Rx - For Prescription Use Only

## D Special Instrument Requirements:

Beckman Coulter AU 680 chemistry analyzer was used to generate data for this submission.

## IV Device/System Characteristics:

### A Device Description:

The ARK Hydrocodone Assay is a homogeneous enzyme immunoassay (HEIA) technique used for the analysis of Hydrocodone in human urine. The ARK Hydrocodone Assay test system includes separately provided test kits for the ARK Hydrocodone Assay.

The ARK Hydrocodone Assay consists of reagents R1 and R2.

Reagent R1 – Antibody/Substrate solution contains: Rabbit monoclonal antibodies to hydrocodone, glucose-6-phosphate, nicotinamide adenine dinucleotide, bovine serum albumin, sodium azide, and stabilizers.

Reagent R2 – Enzyme solution contains: Hydrocodone derivative labeled with recombinant glucose-6-phosphate dehydrogenase (rG6PDH), bovine serum albumin, buffer, sodium azide and stabilizer.

### B Principle of Operation:

The ARK Hydrocodone Assay is a homogeneous enzyme immunoassay. The assay uses specific antibodies that can detect hydrocodone and its metabolites without any significant cross-

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reactivity to other opiate compounds. The assay is based on competition between a drug labeled with recombinant glucose-6-phosphate dehydrogenase (rG6PDH) and free drug from the urine sample, for a fixed amount of specific antibody binding sites. In the absence of free drug from the sample, rabbit monoclonal anti-hydrocodone antibody binds to the drug labeled with rG6PDH and causes a decrease in enzyme activity. In the presence of hydrocodone from the specimen, enzyme activity increases and is directly related to the hydrocodone concentration. Endogenous G6PDH does not interfere because the coenzyme NAD functions only with the bacterial enzyme used in the assay. The enzyme activity is determined spectrophotometrically at 340 nm by measuring the conversion of nicotinamide adenine dinucleotide (NAD) to NADH.

## V Substantial Equivalence Information:

A Predicate Device Name(s): DRI Hydrocodone Assay

B Predicate 510(k) Number(s): K150502

C Comparison with Predicate(s):

|  Device & Predicate Device(s): | K231752 | K150502  |
| --- | --- | --- |
|  Device Trade Name | ARK Hydrocodone Assay | DRI Hydrocodone Assay  |
|  General Device Characteristic Similarities |  |   |
|  Intended Use/Indications For Use | For the detection and estimation of hydrocodone in human urine | Same  |
|  Assay cutoff | 300 ng/mL of Hydrocodone | Same  |
|  Test system type | Homogenous enzyme immunoassay | Same  |
|  Sample Matrix | Human urine | Same  |
|  General Device Characteristic Differences |  |   |
|  Antibody | Rabbit monoclonal | Mouse monoclonal  |
|  Calibrator Levels | 0, 50, 100, 300, 800 ng/mL | 0, 100, 300, 500, 1000 ng/mL  |

## VI Standards/Guidance Documents Referenced:

- CLSI EP12-A2: User Protocol for Evaluation of Qualitative Test Performance.
- CLSI EP05-A3: Evaluation of Precision of Quantitative Measurement Procedures; Approved Guideline - Third Edition.
- CLSI EP07-A3: Interference Testing in Clinical Chemistry.

K231752 - Page 3 of 15

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- CLSI EP37: Supplemental Tables for Interference Testing in Clinical Chemistry - First Edition.

## VII Performance Characteristics (if/when applicable):

## A Analytical Performance:

### 1. Precision/Reproducibility:

A precision study was performed based upon recommendations in the CLSI (EP5-A3) guideline. Drug free urine samples were spiked with hydrocodone at 0, 75, 150, 225, 300, 375, 450, 525 and 600 ng/mL, representing 0, 25, 50, 75, 100, 125, 150, 175 and 200% of the device cutoff (300 ng/mL). Each sample was tested in quadruplicate per run, two runs per day for twenty consecutive days (total N= 160/level/reagent lot) using 2 reagent lots on the Beckman Coulter AU680 chemistry analyzer. The results are shown below:

Qualitative Precision Results:

|  Human Urine (ng/mL) | Relative % Cutoff | # of Results | Lot 1 Precision Results | Lot 2 Precision Results  |
| --- | --- | --- | --- | --- |
|  0 | -100 | 160 | 160 Negative | 160 Negative  |
|  75 | -75 | 160 | 160 Negative | 160 Negative  |
|  150 | -50 | 160 | 160 Negative | 160 Negative  |
|  225 | -25 | 160 | 160 Negative | 160 Negative  |
|  300 | Cutoff | 160 | 50 Negative; 110 Positive | 69 Negative; 91 Positive  |
|  375 | +25 | 160 | 160 Positive | 160 Positive  |
|  450 | +50 | 160 | 160 Positive | 160 Positive  |
|  525 | +75 | 160 | 160 Positive | 160 Positive  |
|  600 | +100 | 160 | 160 Positive | 160 Positive  |

Semi-Quantitative Precision Results:

Semi-quantitative Precision Lot 1

|  Human Urine (ng/mL) | Relative % Cutoff | Precision Results | Repeatability (Within-Run Precision) |   | Between Day Precision |   | Total Precision (Within-Laboratory Precision)  |   |
| --- | --- | --- | --- | --- | --- | --- | --- | --- |
|   |   |   |  SD | CV (%) | SD | CV (%) | SD | CV (%)  |
|  0 | -100 | 160 Negative | 1.2 | N/A | 0.4 | N/A | 1.2 | N/A  |
|  75 | -75 | 160 Negative | 2.3 | 3.0 | 1.8 | 2.3 | 3.0 | 3.9  |
|  150 | -50 | 160 Negative | 6.3 | 4.4 | 4.0 | 2.8 | 7.5 | 5.3  |
|  225 | -25 | 160 Negative | 7.8 | 3.4 | 5.5 | 2.4 | 10.1 | 4.4  |
|  300 | Cutoff | 24 Negative; | 11.1 | 3.5 | 7.9 | 2.5 | 14.7 | 4.7  |

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|  Human Urine (ng/mL) | Relative % Cutoff | Precision Results | Repeatability (Within-Run Precision) |   | Between Day Precision |   | Total Precision (Within-Laboratory Precision)  |   |
| --- | --- | --- | --- | --- | --- | --- | --- | --- |
|   |   |   |  SD | CV (%) | SD | CV (%) | SD | CV (%)  |
|   |   |  136 Positive |  |  |  |  |  |   |
|  375 | +25 | 160 Positive | 14.3 | 3.7 | 11.2 | 2.9 | 18.8 | 4.8  |
|  450 | +50 | 160 Positive | 20.9 | 4.5 | 16.5 | 3.6 | 27.9 | 6.1  |
|  525 | +75 | 160 Positive | 31.7 | 5.9 | 24.2 | 4.5 | 43.6 | 8.1  |
|  600 | +100 | 160 Positive | 50.4 | 8.1 | 38.3 | 6.2 | 61.4 | 9.9  |

Semi-quantitative Precision Lot 2

|  Human Urine (ng/mL) | Relative % Cutoff | Precision Results | Repeatability (Within-Run Precision) |   | Between Day Precision |   | Total Precision (Within-Laboratory Precision)  |   |
| --- | --- | --- | --- | --- | --- | --- | --- | --- |
|   |   |   |  SD | CV (%) | SD | CV (%) | SD | CV (%)  |
|  0 | -100 | 160 Negative | 1.4 | N/A | 0.7 | N/A | 1.6 | N/A  |
|  75 | -75 | 160 Negative | 3.2 | 4.2 | 1.8 | 2.4 | 3.7 | 4.9  |
|  150 | -50 | 160 Negative | 3.3 | 2.3 | 2.1 | 1.4 | 4.5 | 3.1  |
|  225 | -25 | 160 Negative | 6.8 | 2.9 | 4.3 | 1.9 | 8.1 | 3.5  |
|  300 | Cutoff | 35 Negative; 125 Positive | 10.2 | 3.3 | 6.9 | 2.2 | 11.7 | 3.8  |
|  375 | +25 | 160 Positive | 16.7 | 4.3 | 12.0 | 3.1 | 20.0 | 5.2  |
|  450 | +50 | 160 Positive | 26.3 | 5.7 | 11.5 | 2.5 | 27.3 | 5.9  |
|  525 | +75 | 160 Positive | 34.1 | 6.5 | 10.8 | 2.1 | 35.0 | 6.7  |
|  600 | +100 | 160 Positive | 45.6 | 7.5 | 10.1 | 1.7 | 45.6 | 7.5  |

2. Linearity/Recovery:

Linearity study in the semi-quantitative mode was conducted by spiking a drug free urine pool with hydrocodone (serial dilutions of a high concentration hydrocodone in negative urine pool) to achieve concentrations ranging from $0\mathrm{ng / mL}$ to $800\mathrm{ng / mL}$, and testing each level on two reagent lots in replicates of five on the Beckman Coulter AU680 clinical chemistry analyzer. The results from a representative lot are summarized below:

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|  Expected Value (ng/mL) | Observed Value (ng/mL) | Recovery (%)  |
| --- | --- | --- |
|  0 | 0.0 | N/A  |
|  80 | 79.4 | 99.3  |
|  160 | 151.1 | 94.5  |
|  240 | 246.8 | 102.9  |
|  320 | 321.7 | 100.5  |
|  400 | 385.8 | 96.5  |
|  480 | 472.3 | 98.4  |
|  560 | 537.4 | 96.0  |
|  640 | 606.4 | 94.7  |
|  720 | 620.6 | 86.2  |
|  800 | 737.5 | 92.2  |

# 3. Analytical Specificity/Interference:

# Specificity:

An analytical specificity study to evaluate interference from non-structurally and structurally related compounds was performed in the qualitative and semi-quantitative mode. The study design and results are described below. Results were the same for each mode (qualitative and semi-quantitative modes).

# Hydrocodone and its Metabolites

The following metabolites of hydrocodone were approximately equivalent to the  $300\mathrm{ng / mL}$  cutoff at the concentrations tested with the ARK Hydrocodone Assay. Dihydrocodeine tested negative at  $100,000\mathrm{ng / mL}$ . Immunoassay reactivity for hydrocodone and hydromorphone were equivalent.

|  Compound | Concentration Approximately Equivalent to the Cutoff (ng/mL) | Percent Cross-reactivity (%)  |
| --- | --- | --- |
|  Hydrocodone | 292 | 102.8  |
|  Hydromorphone | 299 | 100.4  |
|  Hydromorphone-3β-Glucuronide | 45,439 | 0.7  |
|  Norhydrocodone | 2,277 | 13.2  |
|  Dihydrocodeine | >100,000 | <0.3  |

# Structurally Related Compounds and Other Opiates

To evaluate potential cross-reactivity for the ARK Hydrocodone assay, structurally similar compounds were spiked into drug free urine at concentrations that will yield a result that is equivalent to the  $300\mathrm{ng / mL}$  cutoff. For compounds that did not produce a positive result, the

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highest concentration tested was used to calculate percent cross-reactivity. The percent cross-reactivity is presented in the table below.

|  Compound | Concentration Tested (ng/mL) | Result N=3 (POS/NEG) | Percent Cross-reactivity (%)  |
| --- | --- | --- | --- |
|  6-Acetyl morphine | 100,000 | NEG | <0.3  |
|  Buprenorphine | 100,000 | NEG | <0.3  |
|  Buprenorphine-3β-D-glucuronide | 50,000 | NEG | <0.6  |
|  Codeine | 100,000 | NEG | <0.3  |
|  Codeine-6β-D-glucuronide | 100,000 | NEG | <0.3  |
|  Dextromethorphan | 250,000 | NEG | <0.1  |
|  EDDP | 100,000 | NEG | <0.3  |
|  EMDP | 100,000 | NEG | <0.3  |
|  Ethyl morphine | 100,000 | NEG | <0.3  |
|  Fentanyl | 100,000 | NEG | <0.3  |
|  Heroin | 100,000 | NEG | <0.3  |
|  Levorphanol | 100,000 | NEG | <0.3  |
|  Meperidine | 100,000 | NEG | <0.3  |
|  Methadone | 100,000 | NEG | <0.3  |
|  Morphine | 100,000 | NEG | <0.3  |
|  Morphine-3β-D-glucuronide | 100,000 | NEG | <0.3  |
|  Morphine-6β-D-glucuronide | 100,000 | NEG | <0.3  |
|  Nalbuphine | 100,000 | NEG | <0.3  |
|  Naloxegol | 100,000 | NEG | <0.3  |
|  Naloxone | 100,000 | NEG | <0.3  |
|  Naltrexone | 100,000 | NEG | <0.3  |
|  Norbuprenorphine | 100,000 | NEG | <0.3  |
|  Norcodeine | 100,000 | NEG | <0.3  |
|  Normorphine | 100,000 | NEG | <0.3  |
|  Noroxycodone | 100,000 | NEG | <0.3  |
|  Nortilidine | 100,000 | NEG | <0.3  |
|  Oxycodone | 100,000 | NEG | <0.3  |
|  Oxymorphone | 100,000 | NEG | <0.3  |
|  Oxymorphone-3β-D-glucuronide | 50,000 | NEG | <0.6  |
|  Pentazocine | 100,000 | NEG | <0.3  |
|  Tapentadol | 100,000 | NEG | <0.3  |
|  Thebaine | 100,000 | NEG | <0.3  |
|  Tilidine | 100,000 | NEG | <0.3  |
|  Tramadol | 100,000 | NEG | <0.3  |

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Non-Structurally Related Compounds

Potential interference from non-structurally related drugs and metabolites was evaluated in the qualitative and semi-quantitative modes, by spiking these compounds at high concentrations in drug free urine spiked with hydrocodone at  $\pm 25\%$  of the cutoff (225 and  $375~\mathrm{ng / mL}$ ).

|  Compound | Concentration Tested (ng/mL) | Result N=3 (POS/NEG)  |
| --- | --- | --- |
|  (+)-MDA | 100,000 | NEG  |
|  11-hydroxy-delta-9-THC | 100,000 | NEG  |
|  11-nor-9 carboxy THC | 50,000 | NEG  |
|  1R,2S(-)-Ephedrine | 100,000 | NEG  |
|  1S,2R(+)-Ephedrine | 100,000 | NEG  |
|  4-Bromo-2,5,Dimethoxyphenethylamine | 100,000 | NEG  |
|  7-Aminoclonazepam | 100,000 | NEG  |
|  Acetaminophen | 500,000 | NEG  |
|  Acetylsalicylic acid | 500,000 | NEG  |
|  Alprazolam | 100,000 | NEG  |
|  Amitriptyline | 100,000 | NEG  |
|  Amobarbital | 100,000 | NEG  |
|  Amoxicillin | 100,000 | NEG  |
|  Amphetamine | 100,000 | NEG  |
|  Atorvastatin | 100,000 | NEG  |
|  Benzoylecgonine | 1,000,000 | NEG  |
|  Benzylpiperazine | 100,000 | NEG  |
|  Bupropion | 100,000 | NEG  |
|  Butabarbital | 100,000 | NEG  |
|  Caffeine | 100,000 | NEG  |
|  Canagliflozin | 50,000 | NEG  |
|  Cannabidiol | 100,000 | NEG  |
|  Cannabinol | 100,000 | NEG  |
|  Carbamazepine | 500,000 | NEG  |
|  Carisoprodol | 100,000 | NEG  |
|  Chlordiazepoxide | 100,000 | NEG  |
|  Chlorpromazine | 100,000 | NEG  |
|  Cimetidine | 500,000 | NEG  |
|  Clobazam | 100,000 | NEG  |
|  Clomipramine | 100,000 | NEG  |
|  Clopidogrel | 100,000 | NEG  |
|  Cocaine | 100,000 | NEG  |
|  Cotinine | 100,000 | NEG  |
|  Cyclobenzaprine | 100,000 | NEG  |
|  Desipramine | 100,000 | NEG  |
|  Diazepam | 100,000 | NEG  |

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|  Compound | Concentration Tested (ng/mL) | Result N=3 (POS/NEG)  |
| --- | --- | --- |
|  Diphenhydramine | 100,000 | NEG  |
|  Doxepin | 100,000 | NEG  |
|  Ecgonine | 100,000 | NEG  |
|  Ephedrine | 1,000,000 | NEG  |
|  Fluoxetine | 100,000 | NEG  |
|  Fluphenazine | 100,000 | NEG  |
|  Ibuprofen | 500,000 | NEG  |
|  Imipramine | 100,000 | NEG  |
|  Ketamine | 100,000 | NEG  |
|  Lamotrigine | 100,000 | NEG  |
|  Lidocaine | 100,000 | NEG  |
|  LSD | 100,000 | NEG  |
|  Maprotiline | 100,000 | NEG  |
|  MDMA | 50,000 | NEG  |
|  Meprobamate | 100,000 | NEG  |
|  Metformin | 100,000 | NEG  |
|  Methylphenidate | 250,000 | NEG  |
|  Metronidazole | 100,000 | NEG  |
|  Naproxen | 100,000 | NEG  |
|  Norpseudoephedrine | 50,000 | NEG  |
|  Nortriptyline | 100,000 | NEG  |
|  Omeprazole | 100,000 | NEG  |
|  Ondansetron | 100,000 | NEG  |
|  Oxazepam | 250,000 | NEG  |
|  Phencyclidine | 100,000 | NEG  |
|  Phenobarbital | 100,000 | NEG  |
|  Phentermine | 100,000 | NEG  |
|  Phenylephrine | 100,000 | NEG  |
|  Phenylpropanolamine | 100,000 | NEG  |
|  Phenytoin | 100,000 | NEG  |
|  PMA | 100,000 | NEG  |
|  Propranolol | 100,000 | NEG  |
|  Protriptyline | 100,000 | NEG  |
|  R,R(-)-Pseudoephedrine | 100,000 | NEG  |
|  Ranitidine | 500,000 | NEG  |
|  Ritalinic Acid | 100,000 | NEG  |
|  S(+)-Methamphetamine | 100,000 | NEG  |
|  S,S(+)-Pseudoephedrine | 100,000 | NEG  |
|  Salicylic Acid | 100,000 | NEG  |
|  Secobarbital | 100,000 | NEG  |
|  Sertraline | 100,000 | NEG  |
|  Temazepam | 100,000 | NEG  |

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|  Compound | Concentration Tested (ng/mL) | Result N=3 (POS/NEG)  |
| --- | --- | --- |
|  Theophylline | 50,000 | NEG  |
|  Thioridazine | 100,000 | NEG  |
|  Trazodone | 100,000 | NEG  |
|  Triazolam | 250,000 | NEG  |
|  Trimipramine | 100,000 | NEG  |
|  Venlafaxine | 100,000 | NEG  |
|  Zolpidem | 100,000 | NEG  |

## Endogenous Compounds

Potential interference from endogenous compounds commonly found in urine was evaluated in the qualitative and semi-quantitative modes, by spiking these compounds into drug free urine containing hydrocodone at $\pm 25\%$ of the $300~\mathrm{ng/mL}$ cutoff (225 and $375~\mathrm{ng/mL}$).

|  Interfering Substances | Concentration Tested (mg/dL) | 225 ng/mL Hydrocodone N=3 (POS/NEG) | 375 ng/mL Hydrocodone N=3 (POS/NEG)  |
| --- | --- | --- | --- |
|  Acetaminophen | 10 | NEG | POS  |
|  Acetone | 500 | NEG | POS  |
|  Acetyl Salicylic Acid | 10 | NEG | POS  |
|  Ascorbic acid | 150 | NEG | POS  |
|  Caffeine | 10 | NEG | POS  |
|  Creatinine | 400 | NEG | POS  |
|  Ethanol | 10 | NEG | POS  |
|  Galactose | 5 | NEG | POS  |
|  Glucose | 1000 | NEG | POS  |
|  Hemoglobin | 150 | NEG | POS  |
|  Human Serum Albumin | 200 | NEG | POS  |
|  Human γ-Globulin | 500 | NEG | POS  |
|  Ibuprofen | 10 | NEG | POS  |
|  NaCl | 1000 | NEG | POS  |
|  Oxalic Acid | 50 | NEG | POS  |
|  Riboflavin | 3 | NEG | POS  |
|  Urea | 1000 | NEG | POS  |

## pH and Specific Gravity:

For potential interference from the pH of urine, device performance in the qualitative and semi-quantitative modes was tested using a range of urine pH values (3.0, 4.0, 5.0, 6.0, 7.0, 8.0, 9.0, 10.0 and 11.0). All test samples were prepared in drug free urine containing hydrocodone at $\pm 25\%$ of the cutoff (225 ng/mL and 375 ng/mL hydrocodone

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concentrations). No positive or negative interference was observed at urine pH values ranging from 3.0 to 11.0 for each test mode.

For potential interference from the specific gravity of urine, device performance in the qualitative and semi-quantitative modes was tested using a range of urine specific gravity values (1.000, 1.0021, 1.0043, 1.0179, 1.0187, 1.0262, 1.0303). All test samples were prepared in drug free urine containing hydrocodone at ± 25% of the cutoff (225 ng/mL and 375 ng/mL hydrocodone concentrations). No positive or negative interference was observed at urine specific gravity values ranging from 1.000 to 1.0303 for each test mode.

## Boric Acid:

One percent (1%) w/v of boric acid was tested in the presence of hydrocodone at 225 ng/mL and 375 ng/mL.

|  Compound | Concentration Tested | 225 ng/mL Hydrocodone N=3 (POS/NEG) | 375 ng/mL Hydrocodone N=3 (POS/NEG)  |
| --- | --- | --- | --- |
|  Boric Acid | 1% w/v | NEG | NEG  |

The following limitations statement is included in the Instructions for Use/Package Insert:

“Do not use Boric Acid as a preservative.”

4. Assay Reportable Range:

See the linearity section, VII.A.2., above.

5. Traceability, Stability, Expected Values (Controls, Calibrators, or Methods):

Traceability: ARK Hydrocodone Assay is traceable to a commercially available standard.

6. Detection Limit:

Not applicable.

7. Assay Cut-Off:

Characterization of how the device performs analytically around the claimed cutoff concentration of 300 ng/mL hydrocodone is described in the precision section, VII.A.1., above.

K231752 - Page 11 of 15

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B Comparison Studies:

1. Method Comparison with Predicate Device:

A total of 226 unaltered urine samples from pain management laboratories were analyzed by two lots of the candidate device in the qualitative and semi-quantitative modes on the Beckman Coulter AU680 clinical chemistry analyzer and the comparative mass spectrometry based quantitative method (LC-MS/MS). LC-MS/MS and immunoassay qualitative results are based on a 300ng/mL cutoff. The results from the study in the two modes are summarized below:

Lot 1 Qualitative method comparison with LC-MS/MS as reference method

|  ARK Hydrocodone Assay Results | <50% of cutoff concentration by LC-MS/MS | Near Cutoff Negative (Between 50% below the cutoff and the cutoff concentration by LC-MS/MS) (150-299 ng/mL) | Near Cutoff Positive (Between the cutoff and 50% above the cutoff concentration by LC-MS/MS) (300-450 ng/mL) | High Positive (Greater than 50% above the cutoff concentration by LC-MS/MS)  |
| --- | --- | --- | --- | --- |
|  Positive | 8* | 8* | 9 | 66  |
|  Negative | 134 | 0 | 1* | 0  |

Lot 2 Qualitative method comparison with LC-MS/MS as reference method

|  ARK Hydrocodone Assay Lot 2 Results | <50% of cutoff concentration by LC-MS/MS (<150 ng/mL) | Near Cutoff Negative (Between 50% below the cutoff and the cutoff concentration by LC-MS/MS) (150-299 ng/mL) | Near Cutoff Positive (Between the cutoff and 50% above the cutoff concentration by LC-MS/MS) (300-450 ng/mL) | High Positive (Greater than 50% above the cutoff concentration by LC-MS/MS) (>450 ng/mL)  |
| --- | --- | --- | --- | --- |
|  Positive | 8* | 8* | 9 | 66  |
|  Negative | 134 | 0 | 1* | 0  |

K231752 - Page 12 of 15

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Lot 1 Semi-quantitative method comparison with LC-MS/MS as reference method

|  ARK Hydrocodone Assay Lot 1 Results | <50% of cutoff concentration by LC-MS/MS (<150 ng/mL) | Near Cutoff Negative (Between 50% below the cutoff and the cutoff concentration by LC-MS/MS) (150-299 ng/mL) | Near Cutoff Positive (Between the cutoff and 50% above the cutoff concentration by LC-MS/MS) (300-450 ng/mL) | High Positive (Greater than 50% above the cutoff concentration by LC-MS/MS) (>450 ng/mL)  |
| --- | --- | --- | --- | --- |
|  Positive | 8* | 8* | 9 | 66  |
|  Negative | 134 | 0 | 1* | 0  |

Lot 2 Semi-quantitative method comparison with LC-MS/MS as reference method

|  ARK Hydrocodone Assay Lot 2 Results | <50% of cutoff concentration by LC-MS/MS (<150 ng/mL) | Near Cutoff Negative (Between 50% below the cutoff and the cutoff concentration by LC-MS/MS) (150-299 ng/mL) | Near Cutoff Positive (Between the cutoff and 50% above the cutoff concentration by LC-MS/MS) (300-450 ng/mL) | High Positive (Greater than 50% above the cutoff concentration by LC-MS/MS) (>450 ng/mL)  |
| --- | --- | --- | --- | --- |
|  Positive | 8* | 8* | 10 | 66  |
|  Negative | 134 | 0 | 0 | 0  |

*Discordant Results

|  Sample # | ARK Lot 1 Qual. (POS/NEG) | ARK Lot 2 Qual. (POS/NEG) | LC-MS/MS (ng/mL) |   | ARK Lot 1 Semi-quant. (ng/mL) | ARK Lot 2 Semi-quant. (ng/mL)  |
| --- | --- | --- | --- | --- | --- | --- |
|   |   |   |  Hydrocodone | Hydromorphone |  |   |
|  3 | POS | POS | 287.8 | 210.0 | 476.0 | 664.3  |
|  15 | POS | POS | 226.9 | 150.5 | 390.0 | 415.0  |
|  18 | POS | POS | 156.0 | 174.7 | 335.2 | 338.6  |
|  23 | POS | POS | 5.4 | 1317.7 | 387.7 | 467.9  |
|  38 | NEG | NEG | 306.5 | 22.4 | 277.6 | 307.0  |
|  39 | POS | POS | 162.0 | 52.4 | 316.1 | 312.8  |
|  48 | POS | POS | 200.5 | 61.7 | 358.4 | 347.9  |
|  51 | POS | POS | 174.4 | 29.0 | 357.4 | 393.0  |

K231752 - Page 13 of 15

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|  Sample # | ARK Lot 1 Qual. (POS/NEG) | ARK Lot 2 Qual. (POS/NEG) | LC-MS/MS (ng/mL) |   | ARK Lot 1 Semi-quant. (ng/mL) | ARK Lot 2 Semi-quant. (ng/mL)  |
| --- | --- | --- | --- | --- | --- | --- |
|   |   |   |  Hydrocodone | Hydromorphone |  |   |
|  66 | POS | POS | 146.7 | 190.3 | 463.1 | 440.5  |
|  68 | POS | POS | 181.2 | 150.3 | 382.2 | 376.7  |
|  70 | POS | POS | Not Detected | 545.7 | 445.7 | 480.2  |
|  75 | POS | POS | 5.9 | 10524.1 | 2549.1 | 2922.0  |
|  86 | POS | POS | 255.8 | 30.7 | 471.4 | 412.4  |
|  90 | POS | POS | 106.5 | 214.0 | 335.5 | 333.7  |
|  97 | POS | POS | Not Detected | 1769.8 | 501.3 | 483.9  |
|  99 | POS | POS | Not Detected | 706.1 | 657.9 | 609.1  |
|  101 | POS | POS | Not Detected | 5461.7 | 2014.2 | 2112.0  |

For Sample #38, the hydrocodone concentration was 306.5 ng/mL by LC-MS/MS and 307.0 ng/mL (positive) in semi-quantitative protocol and negative in the qualitative mode. The remaining 16 discordant samples all tested positive, greater than cutoff, by both ARK assay protocols. Cross-reactivity to the major metabolite hydromorphone contributed to the positive results for some of the samples with &lt;300 ng/mL hydrocodone by LC-MS/MS.

2. Matrix Comparison:
Not applicable.

C Clinical Studies:

1. Clinical Sensitivity:
Not applicable.

2. Clinical Specificity:
Not applicable.

3. Other Clinical Supportive Data (When 1. and 2. Are Not Applicable):
Not applicable.

D Clinical Cut-Off:
Not applicable.

K231752 - Page 14 of 15

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E Expected Values/Reference Range:

Not applicable.

VIII Proposed Labeling:

The labeling supports the finding of substantial equivalence for this device.

IX Conclusion:

The submitted information in this premarket notification is complete and supports a substantial equivalence decision.

K231752 - Page 15 of 15

---

**Source:** [https://fda.innolitics.com/submissions/CH/subpart-d%E2%80%94clinical-toxicology-test-systems/DJG/K231752](https://fda.innolitics.com/submissions/CH/subpart-d%E2%80%94clinical-toxicology-test-systems/DJG/K231752)

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